1 #ifndef Mothur_makecontigscommand_h
2 #define Mothur_makecontigscommand_h
5 // makecontigscommand.h
8 // Created by Sarah Westcott on 5/15/12.
9 // Copyright (c) 2012 Schloss Lab. All rights reserved.
12 #include "command.hpp"
13 #include "sequence.hpp"
14 #include "qualityscores.h"
15 #include "alignment.hpp"
16 #include "gotohoverlap.hpp"
17 #include "needlemanoverlap.hpp"
18 #include "blastalign.hpp"
19 #include "noalign.hpp"
20 #include "trimoligos.h"
27 fastqRead() { name = ""; sequence = ""; scores.clear(); };
28 fastqRead(string n, string s, vector<int> sc) : name(n), sequence(s), scores(sc) {};
32 struct pairFastqRead {
37 pairFastqRead(fastqRead f, fastqRead r) : forward(f), reverse(r){};
40 /**************************************************************************************************/
42 class MakeContigsCommand : public Command {
44 MakeContigsCommand(string);
46 ~MakeContigsCommand(){}
48 vector<string> setParameters();
49 string getCommandName() { return "make.contigs"; }
50 string getCommandCategory() { return "Sequence Processing"; }
51 //commmand category choices: Sequence Processing, OTU-Based Approaches, Hypothesis Testing, Phylotype Analysis, General, Clustering and Hidden
53 string getHelpString();
54 string getOutputPattern(string);
55 string getCitation() { return "http://www.mothur.org/wiki/Make.contigs"; }
56 string getDescription() { return "description"; }
59 void help() { m->mothurOut(getHelpString()); }
62 bool abort, allFiles, createGroup;
63 string outputDir, ffastqfile, rfastqfile, align, oligosfile, rfastafile, ffastafile, rqualfile, fqualfile, file;
64 float match, misMatch, gapOpen, gapExtend;
65 int processors, longestBase, threshold, tdiffs, bdiffs, pdiffs, ldiffs, sdiffs;
66 vector<string> outputNames;
68 map<int, oligosPair> barcodes;
69 map<int, oligosPair> primers;
70 vector<string> linker;
71 vector<string> spacer;
72 vector<string> primerNameVector;
73 vector<string> barcodeNameVector;
75 map<string, int> groupCounts;
76 map<string, string> groupMap;
78 fastqRead readFastq(ifstream&, bool&);
79 vector< vector< vector<string> > > preProcessData(unsigned long int&);
80 vector< vector<string> > readFileNames(string);
81 vector< vector<string> > readFastqFiles(unsigned long int&, string, string);
82 vector< vector<string> > readFastaFiles(unsigned long int&, string, string);
83 bool checkReads(fastqRead&, fastqRead&, string, string);
84 int createProcesses(vector< vector<string> >, string, string, string, string, string, vector<vector<string> >, vector<vector<string> >);
85 int driver(vector<string>, string, string, string, string, string, vector<vector<string> >, vector<vector<string> >);
86 bool getOligos(vector<vector<string> >&, vector< vector<string> >&, string);
87 string reverseOligo(string);
88 vector<pairFastqRead> getReads(bool ignoref, bool ignorer, fastqRead forward, fastqRead reverse, map<string, fastqRead>& uniques);
91 /**************************************************************************************************/
93 /**************************************************************************************************/
94 //custom data structure for threads to use.
95 // This is passed by void pointer so it can be any data type
96 // that can be passed using a single void pointer (LPVOID).
100 string outputScrapFasta;
101 string outputScrapQual;
102 string outputMisMatches;
104 vector<string> files;
105 vector<vector<string> > fastaFileNames;
106 vector<vector<string> > qualFileNames;
108 float match, misMatch, gapOpen, gapExtend;
109 int count, threshold, threadID, pdiffs, bdiffs, tdiffs;
110 bool allFiles, createGroup;
111 map<string, int> groupCounts;
112 map<string, string> groupMap;
113 vector<string> primerNameVector;
114 vector<string> barcodeNameVector;
115 map<int, oligosPair> barcodes;
116 map<int, oligosPair> primers;
119 contigsData(vector<string> f, string of, string oq, string osf, string osq, string om, string al, MothurOut* mout, float ma, float misMa, float gapO, float gapE, int thr, map<int, oligosPair> br, map<int, oligosPair> pr, vector<vector<string> > ffn, vector<vector<string> > qfn, vector<string>bnv, vector<string> pnv, int pdf, int bdf, int tdf, bool cg, bool all, int tid) {
123 outputMisMatches = om;
132 outputScrapFasta = osf;
133 outputScrapQual = osq;
134 fastaFileNames = ffn;
138 barcodeNameVector = bnv;
139 primerNameVector = pnv;
149 /**************************************************************************************************/
150 #if defined (__APPLE__) || (__MACH__) || (linux) || (__linux) || (__linux__) || (__unix__) || (__unix)
152 static DWORD WINAPI MyContigsThreadFunction(LPVOID lpParam){
153 contigsData* pDataArray;
154 pDataArray = (contigsData*)lpParam;
157 int longestBase = 1000;
158 Alignment* alignment;
159 if(pDataArray->align == "gotoh") { alignment = new GotohOverlap(pDataArray->gapOpen, pDataArray->gapExtend, pDataArray->match, pDataArray->misMatch, longestBase); }
160 else if(pDataArray->align == "needleman") { alignment = new NeedlemanOverlap(pDataArray->gapOpen, pDataArray->match, pDataArray->misMatch, longestBase); }
163 string thisffastafile = pDataArray->files[0];
164 string thisfqualfile = pDataArray->files[1];
165 string thisrfastafile = pDataArray->files[2];
166 string thisrqualfile = pDataArray->files[3];
168 if (pDataArray->m->debug) { pDataArray->m->mothurOut("[DEBUG]: ffasta = " + thisffastafile + ".\n[DEBUG]: fqual = " + thisfqualfile + ".\n[DEBUG]: rfasta = " + thisrfastafile + ".\n[DEBUG]: rqual = " + thisrqualfile + ".\n"); }
170 if(pDataArray->allFiles){
171 for (int i = 0; i < pDataArray->fastaFileNames.size(); i++) { //clears old file
172 for (int j = 0; j < pDataArray->fastaFileNames[i].size(); j++) { //clears old file
173 if (pDataArray->fastaFileNames[i][j] != "") {
175 pDataArray->m->openOutputFile(pDataArray->fastaFileNames[i][j], temp); temp.close();
176 if (thisfqualfile != "") { pDataArray->m->openOutputFile(pDataArray->qualFileNames[i][j], temp); temp.close(); }
182 ifstream inFFasta, inRFasta, inFQual, inRQual;
183 ofstream outFasta, outQual, outMisMatch, outScrapFasta, outScrapQual;
184 pDataArray->m->openInputFile(thisffastafile, inFFasta);
185 pDataArray->m->openInputFile(thisrfastafile, inRFasta);
186 if (thisfqualfile != "") {
187 pDataArray->m->openInputFile(thisfqualfile, inFQual);
188 pDataArray->m->openInputFile(thisrqualfile, inRQual);
189 pDataArray->m->openOutputFile(pDataArray->outputQual, outQual);
190 pDataArray->m->openOutputFile(pDataArray->outputScrapQual, outScrapQual);
192 pDataArray->m->openOutputFile(pDataArray->outputFasta, outFasta);
193 pDataArray->m->openOutputFile(pDataArray->outputMisMatches, outMisMatch);
194 pDataArray->m->openOutputFile(pDataArray->outputScrapFasta, outScrapFasta);
196 outMisMatch << "Name\tLength\tMisMatches\n";
198 TrimOligos trimOligos(pDataArray->pdiffs, pDataArray->bdiffs, 0, 0, pDataArray->primers, pDataArray->barcodes);
200 while ((!inFFasta.eof()) && (!inRFasta.eof())) {
202 if (pDataArray->m->control_pressed) { break; }
205 string trashCode = "";
206 int currentSeqsDiffs = 0;
208 //read seqs and quality info
209 Sequence fSeq(inFFasta); pDataArray->m->gobble(inFFasta);
210 Sequence rSeq(inRFasta); pDataArray->m->gobble(inRFasta);
211 QualityScores* fQual = NULL; QualityScores* rQual = NULL;
212 if (thisfqualfile != "") {
213 fQual = new QualityScores(inFQual); pDataArray->m->gobble(inFQual);
214 rQual = new QualityScores(inRQual); pDataArray->m->gobble(inRQual);
217 int barcodeIndex = 0;
220 if(pDataArray->barcodes.size() != 0){
221 if (thisfqualfile != "") {
222 success = trimOligos.stripBarcode(fSeq, rSeq, *fQual, *rQual, barcodeIndex);
224 success = trimOligos.stripBarcode(fSeq, rSeq, barcodeIndex);
226 if(success > pDataArray->bdiffs) { trashCode += 'b'; }
227 else{ currentSeqsDiffs += success; }
230 if(pDataArray->primers.size() != 0){
231 if (thisfqualfile != "") {
232 success = trimOligos.stripForward(fSeq, rSeq, *fQual, *rQual, primerIndex);
234 success = trimOligos.stripForward(fSeq, rSeq, primerIndex);
236 if(success > pDataArray->pdiffs) { trashCode += 'f'; }
237 else{ currentSeqsDiffs += success; }
240 if (currentSeqsDiffs > pDataArray->tdiffs) { trashCode += 't'; }
242 //flip the reverse reads
243 rSeq.reverseComplement();
244 if (thisfqualfile != "") { rQual->flipQScores(); }
247 alignment->align(fSeq.getUnaligned(), rSeq.getUnaligned());
248 map<int, int> ABaseMap = alignment->getSeqAAlnBaseMap();
249 map<int, int> BBaseMap = alignment->getSeqBAlnBaseMap();
250 fSeq.setAligned(alignment->getSeqAAln());
251 rSeq.setAligned(alignment->getSeqBAln());
252 int length = fSeq.getAligned().length();
254 //traverse alignments merging into one contiguous seq
256 vector<int> contigScores;
257 int numMismatches = 0;
258 string seq1 = fSeq.getAligned();
259 string seq2 = rSeq.getAligned();
260 vector<int> scores1, scores2;
261 if (thisfqualfile != "") {
262 scores1 = fQual->getQualityScores();
263 scores2 = rQual->getQualityScores();
264 delete fQual; delete rQual;
267 int overlapStart = fSeq.getStartPos();
268 int seq2Start = rSeq.getStartPos();
269 //bigger of the 2 starting positions is the location of the overlapping start
270 if (overlapStart < seq2Start) { //seq2 starts later so take from 0 to seq2Start from seq1
271 overlapStart = seq2Start;
272 for (int i = 0; i < overlapStart; i++) {
274 if (thisfqualfile != "") { contigScores.push_back(scores1[ABaseMap[i]]); }
276 }else { //seq1 starts later so take from 0 to overlapStart from seq2
277 for (int i = 0; i < overlapStart; i++) {
279 if (thisfqualfile != "") { contigScores.push_back(scores2[BBaseMap[i]]); }
283 int seq1End = fSeq.getEndPos();
284 int seq2End = rSeq.getEndPos();
285 int overlapEnd = seq1End;
286 if (seq2End < overlapEnd) { overlapEnd = seq2End; } //smallest end position is where overlapping ends
288 for (int i = overlapStart; i < overlapEnd; i++) {
289 if (seq1[i] == seq2[i]) { //match, add base and choose highest score
291 if (thisfqualfile != "") {
292 contigScores.push_back(scores1[ABaseMap[i]]);
293 if (scores1[ABaseMap[i]] < scores2[BBaseMap[i]]) { contigScores[contigScores.size()-1] = scores2[BBaseMap[i]]; }
295 }else if (((seq1[i] == '.') || (seq1[i] == '-')) && ((seq2[i] != '-') && (seq2[i] != '.'))) { //seq1 is a gap and seq2 is a base, choose seq2, unless quality score for base is below threshold. In that case eliminate base
296 if (thisfqualfile != "") {
297 if (scores2[BBaseMap[i]] < pDataArray->threshold) { } //
300 contigScores.push_back(scores2[BBaseMap[i]]);
302 }else { contig += seq2[i]; }
303 }else if (((seq2[i] == '.') || (seq2[i] == '-')) && ((seq1[i] != '-') && (seq1[i] != '.'))) { //seq2 is a gap and seq1 is a base, choose seq1, unless quality score for base is below threshold. In that case eliminate base
304 if (thisfqualfile != "") {
305 if (scores1[ABaseMap[i]] < pDataArray->threshold) { } //
308 contigScores.push_back(scores1[ABaseMap[i]]);
310 }else { contig += seq1[i]; }
311 }else if (((seq1[i] != '-') && (seq1[i] != '.')) && ((seq2[i] != '-') && (seq2[i] != '.'))) { //both bases choose one with better quality
312 if (thisfqualfile != "") {
314 contigScores.push_back(scores1[ABaseMap[i]]);
315 if (scores1[ABaseMap[i]] < scores2[BBaseMap[i]]) { contigScores[contigScores.size()-1] = scores2[BBaseMap[i]]; c = seq2[i]; }
318 }else { numMismatches++; }
319 }else { //should never get here
320 pDataArray->m->mothurOut("[ERROR]: case I didn't think of seq1 = " + toString(seq1[i]) + " and seq2 = " + toString(seq2[i]) + "\n");
324 if (seq1End < seq2End) { //seq1 ends before seq2 so take from overlap to length from seq2
325 for (int i = overlapEnd; i < length; i++) {
327 if (thisfqualfile != "") { contigScores.push_back(scores2[BBaseMap[i]]); }
329 }else { //seq2 ends before seq1 so take from overlap to length from seq1
330 for (int i = overlapEnd; i < length; i++) {
332 if (thisfqualfile != "") { contigScores.push_back(scores1[ABaseMap[i]]); }
337 if(trashCode.length() == 0){
338 if (pDataArray->createGroup) {
339 if(pDataArray->barcodes.size() != 0){
340 string thisGroup = pDataArray->barcodeNameVector[barcodeIndex];
341 if (pDataArray->primers.size() != 0) {
342 if (pDataArray->primerNameVector[primerIndex] != "") {
343 if(thisGroup != "") {
344 thisGroup += "." + pDataArray->primerNameVector[primerIndex];
346 thisGroup = pDataArray->primerNameVector[primerIndex];
351 if (pDataArray->m->debug) { pDataArray->m->mothurOut(", group= " + thisGroup + "\n"); }
353 pDataArray->groupMap[fSeq.getName()] = thisGroup;
355 map<string, int>::iterator it = pDataArray->groupCounts.find(thisGroup);
356 if (it == pDataArray->groupCounts.end()) { pDataArray->groupCounts[thisGroup] = 1; }
357 else { pDataArray->groupCounts[it->first] ++; }
362 if(pDataArray->allFiles){
364 pDataArray->m->openOutputFileAppend(pDataArray->fastaFileNames[barcodeIndex][primerIndex], output);
365 output << ">" << fSeq.getName() << endl << contig << endl;
368 if (thisfqualfile != "") {
369 pDataArray->m->openOutputFileAppend(pDataArray->qualFileNames[barcodeIndex][primerIndex], output);
370 output << ">" << fSeq.getName() << endl;
371 for (int i = 0; i < contigScores.size(); i++) { output << contigScores[i] << ' '; }
378 outFasta << ">" << fSeq.getName() << endl << contig << endl;
379 if (thisfqualfile != "") {
380 outQual << ">" << fSeq.getName() << endl;
381 for (int i = 0; i < contigScores.size(); i++) { outQual << contigScores[i] << ' '; }
384 outMisMatch << fSeq.getName() << '\t' << contig.length() << '\t' << numMismatches << endl;
387 outScrapFasta << ">" << fSeq.getName() << " | " << trashCode << endl << contig << endl;
388 if (thisfqualfile != "") {
389 outScrapQual << ">" << fSeq.getName() << " | " << trashCode << endl;
390 for (int i = 0; i < contigScores.size(); i++) { outScrapQual << contigScores[i] << ' '; }
391 outScrapQual << endl;
397 if((num) % 1000 == 0){ pDataArray->m->mothurOut(toString(num)); pDataArray->m->mothurOutEndLine(); }
401 if((num) % 1000 != 0){ pDataArray->m->mothurOut(toString(num)); pDataArray->m->mothurOutEndLine(); }
407 outScrapFasta.close();
408 if (thisfqualfile != "") {
412 outScrapQual.close();
416 if (pDataArray->m->control_pressed) { pDataArray->m->mothurRemove(pDataArray->outputFasta); pDataArray->m->mothurRemove(pDataArray->outputMisMatches); pDataArray->m->mothurRemove(pDataArray->outputScrapFasta); if (thisfqualfile != "") { pDataArray->m->mothurRemove(pDataArray->outputQual); pDataArray->m->mothurRemove(pDataArray->outputScrapQual); } }
421 catch(exception& e) {
422 pDataArray->m->errorOut(e, "AlignCommand", "MyContigsThreadFunction");